中国中药杂志

2012, v.37(03) 298-301

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蜈蚣胃蛋白酶解物体外抗凝活性肽类的分离与分析
Study on separation and analysis of anticoagulant compounds for anticoagulant activity in vitro on mixture of peptide from pepsin enzymolysis of centipede

姜丽;王玉蓉;赵韶华;黄能听;
JIANG Li1,WANG Yurong1,ZHAO Shaohua2,HUANG Nengting1(1.School of Chinese Materia Medica,Beijing University of Traditional Chinese Medicine,Beijing 100102,China; 2.Hebei Yiling Pharmaceutical Group,Shijiazhuang 050035,China)

摘要(Abstract):

目的:采用凝胶过滤色谱和C18反相色谱对蜈蚣胃蛋白酶解物的抗凝活性部位进行分离纯化,并检测其相对分子质量分布范围。方法:按色带及280 nm紫外光谱法收集凝胶色谱分离各流分;以活化部位凝血活酶时间(APTT)为指标,检测各流分体外抗凝活性;用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)检测其相对分子质量分布范围。结果:经凝胶过滤色谱和C18反相色谱纯化得到了抗凝活性组分,用MALDI-TOF-MS测得其相对分子质量分布范围为597~1 146。结论:凝胶过滤色谱和C18反相色谱可用于蜈蚣胃蛋白酶解物抗凝活性组分的分离纯化,其抗凝活性组分为小分子肽类。
Objective: To separate anticoagulant components from the pepsin enzymolysis of centipede by gel filtration and reverse-phase C18 chromatography,and to detect the distribution range of their molecular mass.Method: Cingula and 280 nm ultraviolet spectrometry were used to detect and collect the chromatographic solutions.The components′ anticoagulant activity in vitro was detected with activated partial thromboplastin time(APTT) as the index,and the molecular mass range of the active composition was detected by MALDI-TOF-MS.Result: Anticoagulant active compounds were produced by gel filtration and reverse-phase C18 chromatography.The distribution range of relative molecular mass was determined to be from 597 to 1 146.Conclusion: Gel filtration and reverse-phase C18 chromatography are feasible for separating and purifying the pepsin enzymolysis of Centipede.The anticoagulant active compounds are oligopeptides.

关键词(KeyWords): 蜈蚣胃蛋白酶解物;凝胶过滤色谱;C18反相色谱;抗凝;MALDI-TOF-MS
pepsin enzymolysis of centipede;gel filtration chromatography;reverse-phase C18 chromatography;anticoagulant effect;MALDI-TOF-MS

Abstract:

Keywords:

基金项目(Foundation): 国家“重大新药创制”科技重大专项(2009ZX09313-003);; 北京中医药大学复方中药制剂研究创新团队项目(2011-CXTD-13)

作者(Authors): 姜丽;王玉蓉;赵韶华;黄能听;
JIANG Li1,WANG Yurong1,ZHAO Shaohua2,HUANG Nengting1(1.School of Chinese Materia Medica,Beijing University of Traditional Chinese Medicine,Beijing 100102,China; 2.Hebei Yiling Pharmaceutical Group,Shijiazhuang 050035,China)

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